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  • 1
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    New York, NY : Springer New York
    Keywords: Life sciences ; Cell Biology ; Life sciences ; Cell Biology ; Springer eBooks
    Description / Table of Contents: Whole Genome Sequencing-based Discovery of Structural Variants in Glioblastoma -- Whole Genome DNA Methylation Analysis of Human Glioblastoma using Illumina Bead Arrays -- Establishing Primary Human Glioblastoma Adherent Cultures from Operative Specimens -- Establishing Primary Human Glioblastoma Tumorsphere Cultures from Operative Specimens -- Isolation of GBM Stem Cells with Flow Cytometry -- Lentiviral Transduction of Primary Human Glioblastoma Cultures -- Selective Targeting of CD133-expressing Glioblastoma Stem Cells using Lentiviral Vectors -- Intracellular pH Measurements in Glioblastoma Cells using the pH-sensitive Dye BCECF -- Induction and Assessment of Hypoxia in Glioblastoma Cells in vitro -- Metabolomic Analysis of Glioma Cells Using Nanoflow Liquid Chromatography-tandem Mass Spectrometry -- Sample Preparation for Relative Quantitation of Proteins Using Tandem Mass Tags (TMT) and Mass Spectrometry (MS) -- Single-Cell RNA Sequencing of Glioblastoma Cells -- Evaluation of Radioresponse and Radiosensitizers in Glioblastoma Organotypic Cultures -- Orthotopic Patient-derived Glioblastoma Xenografts in Mice -- Bioluminescent In Vivo Imaging of Orthotopic Glioblastoma Xenografts in Mice -- Evaluation of Vascularity, Blood Perfusion, and Oxygen Tension in Tumor Xenografts with Fluorescent Microscopy -- Probing Glioblastoma Tissue Heterogeneity with Laser Capture Microdissection -- Flow Cytometric Identification of Tumor-infiltrating Lymphocytes from Glioblastoma -- Modeling Glioma with Human Embryonic Stem Cell-derived Neural Lineages.℗
    Abstract: This volume details widely used and newer lab℗ protocols for studying hypoxic responses in℗ physiology and diseases. Chapters guide the℗ reader through the application of hypoxic℗ conditions, to the techniques of molecular℗ biology, biochemical, cell biology, genomic,bioinformatic, metabolic, and animal℗ studies. Written in the highly℗ successfulMethods in Molecular Biology℗ series℗ format, chapters include introductions to their℗ respective topics, lists of the necessary℗ materials and reagents, step-by-step, readily℗ reproducible laboratory protocols, and tips on℗ troubleshooting and avoiding known pitfalls.℗ Authoritative and cutting-edge,℗ Hypoxia: Methods℗ and Protocols℗ aims to provide a valuable set of℗ tools that can be used to study hypoxia and℗ beyond
    Pages: XII, 302 p. 73 illus., 63 illus. in color. : online resource.
    ISBN: 9781493976652
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  • 2
    Publication Date: 2018-12-11
    Description: Women are increasingly using botanical dietary supplements (BDS) to reduce menopausal hot flashes. Although licorice ( Glycyrrhiza sp.) is one of the frequently used ingredients in BDS, the exact plant species is often not identified. We previously showed that in breast epithelial cells (MCF-10A), Glycyrrhiza glabra (GG) and G. inflata (GI), and their compounds differentially modulated P450 1A1 and P450 1B1 gene expression, which are responsible for estrogen detoxification and genotoxicity, respectively. GG and isoliquiritigenin (LigC) increased CYP1A1 , whereas GI and its marker compound, licochalcone A (LicA), decreased CYP1A1 and CYP1B1 . The objective of this study was to determine the distribution of the bioactive licorice compounds, the metabolism of LicA, and whether GG, GI, and/or pure LicA modulate NAD(P)H quinone oxidoreductase (NQO1) in an ACI rat model. In addition, the effect of licorice extracts and compounds on biomarkers of estrogen chemoprevention ( CYP1A1 ) as well as carcinogenesis ( CYP1B1 ) was studied. LicA was extensively glucuronidated and formed GSH adducts; however, free LicA as well as LigC were bioavailable in target tissues after oral intake of licorice extracts. GG, GI, and LicA caused induction of NQO1 activity in the liver. In mammary tissue, GI increased CYP1A1 and decreased CYP1B1 , whereas GG only increased CYP1A1 . LigC may have contributed to the upregulation of CYP1A1 after GG and GI administration. In contrast, LicA was responsible for GI-mediated downregulation of CYP1B1 . These studies highlight the polypharmacologic nature of botanicals and the importance of standardization of licorice BDS to specific Glycyrrhiza species and to multiple constituents.
    Print ISSN: 1940-6207
    Electronic ISSN: 1940-6215
    Topics: Medicine
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  • 3
    Publication Date: 2018-11-16
    Description: Purpose: Current tumor–node–metastasis (TNM) staging system cannot provide adequate information for prediction of prognosis and chemotherapeutic benefits. We constructed a classifier to predict prognosis and identify a subset of patients who can benefit from adjuvant chemotherapy. Experimental Design: We detected expression of 15 immunohistochemistry (IHC) features in tumors from 251 gastric cancer (GC) patients and evaluated the association of their expression level with overall survival (OS) and disease-free survival (DFS). Then, integrating multiple clinicopathologic features and IHC features, we used support vector machine (SVM)–based methods to develop a prognostic classifier (GC-SVM classifier) with features. Further validation of the GC-SVM classifier was performed in two validation cohorts of 535 patients. Results: The GC-SVM classifier integrated patient sex, carcinoembryonic antigen, lymph node metastasis, and the protein expression level of eight features, including CD3 invasive margin (IM) , CD3 center of tumor (CT) , CD8 IM , CD45RO CT , CD57 IM , CD66b IM , CD68 CT , and CD34. Significant differences were found between the high- and low-GC-SVM patients in 5-year OS and DFS in training and validation cohorts. Multivariate analysis revealed that the GC-SVM classifier was an independent prognostic factor. The classifier had higher predictive accuracy for OS and DFS than TNM stage and can complement the prognostic value of the TNM staging system. Further analysis revealed that stage II and III GC patients with high-GC-SVM were likely to benefit from adjuvant chemotherapy. Conclusions: The newly developed GC-SVM classifier was a powerful predictor of OS and DFS. Moreover, the GC-SVM classifier could predict which patients with stage II and III GC benefit from adjuvant chemotherapy. Clin Cancer Res; 24(22); 5574–84. ©2018 AACR .
    Print ISSN: 1078-0432
    Electronic ISSN: 1557-3265
    Topics: Medicine
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  • 4
    Publication Date: 2018-04-04
    Description: Purpose: Since drug responses vary between patients, it is crucial to develop pre-clinical or co-clinical strategies that forecast patient response. In this study, we tested whether RNA-based therapeutics were suitable for personalized medicine by using patient-derived-organoid (PDO) and patient-derived-xenograft (PDX) models. Experimental Design: We performed microRNA (miRNA) profiling of PDX samples to determine the status of miRNA deregulation in individual pancreatic ductal adenocarcinoma (PDAC) patients. To deliver personalized RNA-based-therapy targeting oncogenic miRNAs that form part of this common PDAC miRNA over-expression signature, we packaged antimiR oligonucleotides against one of these miRNAs in tumor-penetrating nanocomplexes (TPN) targeting cell surface proteins on PDAC tumors. Results: As a validation for our pre-clinical strategy, the therapeutic potential of one of our nano-drugs, TPN-21, was first shown to decrease tumor cell growth and survival in PDO avatars for individual patients, then in their PDX avatars. Conclusions: This general approach appears suitable for co-clinical validation of personalized RNA medicine and paves the way to prospectively identify patients with eligible miRNA profiles for personalized RNA-based therapy. Clin Cancer Res; 24(7); 1734–47. ©2018 AACR .
    Print ISSN: 1078-0432
    Electronic ISSN: 1557-3265
    Topics: Medicine
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  • 5
  • 6
    Abstract: Astrocytic brain tumours, including glioblastomas, are incurable neoplasms characterized by diffusely infiltrative growth. Here we show that many tumour cells in astrocytomas extend ultra-long membrane protrusions, and use these distinct tumour microtubes as routes for brain invasion, proliferation, and to interconnect over long distances. The resulting network allows multicellular communication through microtube-associated gap junctions. When damage to the network occurred, tumour microtubes were used for repair. Moreover, the microtube-connected astrocytoma cells, but not those remaining unconnected throughout tumour progression, were protected from cell death inflicted by radiotherapy. The neuronal growth-associated protein 43 was important for microtube formation and function, and drove microtube-dependent tumour cell invasion, proliferation, interconnection, and radioresistance. Oligodendroglial brain tumours were deficient in this mechanism. In summary, astrocytomas can develop functional multicellular network structures. Disconnection of astrocytoma cells by targeting their tumour microtubes emerges as a new principle to reduce the treatment resistance of this disease.
    Type of Publication: Journal article published
    PubMed ID: 26536111
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  • 7
  • 8
    Abstract: BACKGROUND & AIMS: DNA ploidy, a DNA flow cytometry parameter, reflects tumor cell cycle. In breast cancer (BC), ploidy status characterizes genotypic stability and potential metastatic capacity. It is suggested that aneuploidy is an independent prognosticator for BC patients and could aid for individualized medicine. There are extensive studies concerning the prognostic significance of DNA aneuploidy, however, its clinical utility remains controversial. Herein we conducted a meta-analysis to determine the correlation between DNA ploidy status and BC characteristics and survival. METHODS: The electronic databases PubMed, EMBASE, and Web of Science were searched for relevant studies. The major investigated parameters were the BC aneuploidy rates in relation to tumor stage, size, lymph node metastasis, grading, estrogen receptor (ER) status, disease-free survival (DFS), and overall survival (OS). Hazard ratios (HRs) and the corresponding 95% confidence intervals (CIs) for DFS and OS were extracted from each study before meta-analyzed. Risk ratios (RRs) were computed using the fixed-effect or random-effects model according to data heterogeneity, and the Mantel-Haenszel or the inverse-variance method was adopted where appropriate to obtain pooled estimates using RevMan 5.3. The Egger's test was conducted with Stata 11. RESULTS: Pooled analyses of data from 29 studies involving a total of 141,163 cases showed that BC patients with more advanced tumors (stage I vs. stages II-IV, RR=0.84; 95% CI, 0.74 to 0.96; P=0.01), larger tumors (〈/=2 cm vs. 〉2 cm: RR=0.82; 95% CI, 0.77 to 0.87; P〈0.00001), lymph node metastasis (pN0 vs. pN1-3: RR=0.85; 95% CI, 0.83 to 0.87, P〈0.00001), poorer tumor proliferation (G2 vs. G1: RR=1.58; 95% CI, 1.40 to 1.79; P〈0.00001; G3 vs. G1: RR=2.17; 95% CI, 1.77 to 2.67; P〈0.00001; G3 vs. G2: RR=1.41; 95% CI, 1.25 to 1.60; P〈0.00001), and ER- status (ER-vs. ER+: RR=1.32; 95% CI, 1.22 to 1.43; P〈0.00001) were significantly more frequently aneuploid. BC patients with diploid tumors had better clinical outcomes than those with aneuploid cancers. The pooled HR estimates were0.73 (P〈0.0001) for DFS and 0.72 (P=0.0001) for OS, respectively. CONCLUSION: This meta-analysis implies that DNA aneuploidy is a significant predictor for BC progression and survival, and should be focused on in the therapeutic planning.
    Type of Publication: Journal article published
    PubMed ID: 27528028
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  • 9
    Abstract: Purpose: Elevated levels of neutrophils have been associated with poor survival in various cancers, but direct evidence supporting a role for neutrophils in the immunopathogenesis of human cancers is lacking.Experimental Design: A total of 573 patients with gastric cancer were enrolled in this study. Immunohistochemistry and real-time PCR were performed to analyze the distribution and clinical relevance of neutrophils in different microanatomic regions. The regulation and function of neutrophils were assessed both in vitro and in vivoResults: Increased neutrophil counts in the peripheral blood were associated with poor prognosis in gastric cancer patients. In gastric cancer tissues, neutrophils were enriched predominantly in the invasive margin, and neutrophil levels were a powerful predictor of poor survival in patients with gastric cancer. IL17+ neutrophils constitute a large portion of IL17-producing cells in human gastric cancer. Proinflammatory IL17 is a critical mediator of the recruitment of neutrophils into the invasive margin by CXC chemokines. Moreover, neutrophils at the invasive margin were a major source of matrix metalloproteinase-9, a secreted protein that stimulates proangiogenic activity in gastric cancer cells. Accordingly, high levels of infiltrated neutrophils at the invasive margin were positively correlated with angiogenesis progression in patients with gastric cancer.Conclusions: These data provide direct evidence supporting the pivotal role of neutrophils in gastric cancer progression and reveal a novel immune escape mechanism involving fine-tuned collaborative action between cancer cells and immune cells in the distinct tumor microenvironment. Clin Cancer Res; 23(6); 1575-85. (c)2016 AACR.
    Type of Publication: Journal article published
    PubMed ID: 27620275
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  • 10
    Keywords: DISEASE ; UNITED-STATES ; JAPANESE POPULATION ; German ; SEQUENCE VARIANTS ; LOXL1 GENE POLYMORPHISMS ; PSEUDOEXFOLIATION SYNDROME ; GLAUCOMA
    Abstract: Exfoliation syndrome (XFS) is the most common recognizable cause of open-angle glaucoma worldwide. To better understand the etiology of XFS, we conducted a genome-wide association study (GWAS) of 1,484 cases and 1,188 controls from Japan and followed up the most significant findings in a further 6,901 cases and 20,727 controls from 17 countries across 6 continents. We discovered a genome-wide significant association between a new locus (CACNA1A rs4926244) and increased susceptibility to XFS (odds ratio (OR) = 1.16, P = 3.36 x 10(-11)). Although we also confirmed overwhelming association at the LOXL1 locus, the key SNP marker (LOXL1 rs4886776) demonstrated allelic reversal depending on the ancestry group (Japanese: ORA allele = 9.87, P = 2.13 x 10(-217); non-Japanese: ORA allele = 0.49, P = 2.35 x 10(-31)). Our findings represent the first genetic locus outside of LOXL1 surpassing genome-wide significance for XFS and provide insight into the biology and pathogenesis of the disease.
    Type of Publication: Journal article published
    PubMed ID: 25706626
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